中国壮族群体人类白细胞抗原HLA-Cw基因遗传多态性的测序分型研究

本研究探讨壮族人群HLA—Cw基因的遗传多态性。采用自行建立的HLA—Cw基因测序分型方法，对150份壮族非血缘个体血样HLA-Cw基因的第2、3、4外显子进行序列测定；测序反应产物用ABIPrism^TM3730测序仪电泳检测，用Assign3．5分析软件分析结果。结果表明：经Assign3．5软件分析，能直接分析出明确的HLA—Cw基因型的样本占33．33％；排除罕见等位基因后，可判定HLA—Cw基因型的样本占63．33％；其余的5份样本（3．33％）经PCR—SSP高分辨分型试剂盒检测后，可判定基因型；共检出了16种HIA．Cw等位基因，频率大于10％的4种常见等位基因为Cw＊0304〉Cw＊0102〉Cw＊0801〉Cw＊0702，基因多样性（genediversity，GD）为0．9297。Cw＊01，03，07，08，12，14（Cw1组）与Cw＊02，04，05，06，15，16，17，18（Cw2组）的频率分别为0．8967和0．1032，与KIR的识别方式以Cw1组等位基因为主。在检出的51种基因型中，纯合子比例占9．33％（14／150），基因型频率的分布符合Hardy．Weinberg定律。结论：本研究所得到的壮族群体HLA—Cw位点的等位基因频率及其分布特点，可为人类学、HIA-Cw基因与疾病关联等研究提供基础数据。

Genetic Polymorphism of Chinese Zhuang Population at HLA-Cw Locus by Sequence Based Typing

Thist study was purposed to explore the genetic polymorphism of Chinese Zhuang population at HLA-Cw locus by sequence based typing （SBT）. A total of 150 unrelated blood samples from Chinese Zhuang population were subjected to sequencing at exon 2, 3 and 4 of HLA-Cw gene in both directions by using SBT technique established by our laboratory. The purified products of sequencing reaction were run by means of electrophoresis on the ABI 3730 DNA Sequencer and the assignment of HLA-Cw genotype was accomplished by using the Assign 3.5 software. The consensus sequence at exon 2, 3 and 4 of HLA-Cw gene for each sample was imported into the Assign 3.5 software. The results showed that 33.33% of tested samples could obtain an unique genotype, genotype in 63.33% of tested samples with ambiguous results could be assigned by ruling out the rare alleles according to the NMDP Rare Allele List File; however, the final genotype in rest 3.33 % of the detected samples could be defined when subjected to further confirmatory testing by PCR-SSP. In this detection 16 HLA-Cw alleles were identified, the common alleles with a frequency of 〉 10% were Cw ＊ 0304 〉 Cw ＊ 0102 〉 Cw ＊ 0801 〉 Cw ＊ 0702. The value for gene diversity （GD） was 0.9297, The frequency for Cw ＊ 01, 03, 07, 08, 12, 14 （Cw 1 allele group） and Cw ＊ 02, 04, 05, 06, 15, 16, 17, 18 （Cw 2 allele group） was 0. 8967 and 0. 1032, respectively,which indicated that the Cw 1 allele group is the dominant ligand for KIR in Chinese Zhuang population. 51 genotypes were determined and the distribution of genotype frequency was in line with Hardy- Weinberg principle. It is concluded that the obtained HLA-Cw allele frequency and its distribution characteristics of Chinese Zhuang population can provide valuable data in the studies of anthropology and the association of HLA-Cw with disease.